Method and preparation for treatment of carcinoma

ABSTRACT

A BACTERIA LYSING AOLUTION IS PRESENTED PREPARED BY INCUBATING BLOOD CULTURES OF STAPHYLOCOCCUS ALBUS CULTURED FROM CARCINOMA TISSUE IN A MULTIPLE ELECTROLYTE INCLUDING 10 PERCENT INVERT SUGAR, AND INSULIN AT BETWEEN 98*-100* F. FOR APPROXIMATELY EIGHT WEEKS. THE SOLUTION IS USEFUL IN TREATING METASTATIC CARCINOMA. PATIENTS SUFFERING WITH CARCINOMA ARE INJECTED INTRAMSCULARLY WITH ABOUT 2 CC. OF THE SOLUTION PER INJECTION. TREATMENT IS REPEATED DAILY FOR FOUR OR FIVE DAYS, AND THEN EVERY TWO OR THREE DAYS FOR FROM ONE TO THREE MONTHS. OBSERVED RESULTS WITH PATIENTS SUFFERING FROM LESS ADVANCED TO METASTATIC CARCINOMA INCLUDE TUMOR REGRESSION, AND ELIMINATION OF PAIN ASSOCIATED WITH THE DISEASE. RESULTS WITH PATIENTS SUFFERING FROM ADVANCED TERMINAL CONDITIONS SHOW COMPLETE ELIMINATION OF PAIN. IN SOME INSTANCES, A COMPLETE TUMOR REGRESSION WAS OBSERVED WITH SUBSEQUENT RETURN TO HEALTH.

United States Patent 3,655,873 METHOD AND PREPARATION FOR TREATMENT OF CARCINOMA Francis M. Duffy, 211 W. Maple, Enid, Okla. 73701 No Drawing. Filed July 24, 1970, Ser. No. 58,204

Int. Cl. C12k 7/00 US. Cl. 424-93 7 Claims ABSTRACT OF THE DISCLOSURE A bacteria lysing solution is presented prepared by incubating blood cultures of Staphylococcus albus cultured from carcinoma tissue in a multiple electrolyte including 10 percent invert sugar, and insulin at between 98100 F. for approximately eight weeks. The solution is useful in treating metastatic carcinoma. Patients suffering with carcinoma are injected intramuscularly with about 2 cc. of the solution per injection. Treatment is repeated daily for four or five days, and then every two to three days for from one to three months. Observed results with patients suffering from less advanced to metastatic carcinoma include tumor regression, and elimination of pain associated with the disease. Results with patients suffering from advanced terminal conditions show complete elimination of pain. In some instances, a complete tumor regression was observed with subsequent return to health.

This invention relates to a preparation and method for therapeutic administration thereof to repress and treat malignant tumors and alleviate pain associated with such conditions in humans. The invention is particularly directed to utilization by persons suffering from metastatic carcinoma. The method of treatment of this invention includes the preparation of a solution which, when injected in patients suffering from carcinoma will result in repression of the tumor growth or metastasis and relief from pain associated with the disease.

Malignancies are generally classified according to the tissue of origin, and are characterized by the properties of anaplasia, invasion, and metastasis. These properties reflect the independence of malignant cells, and their ability to grow at sites other than the locus of origin. Invasion precedes metastasis and tumor progression, when individual or small groups of cells, no longer dependent upon their site of origin for survival, are capable of infiltrating the surrounding host tissue and remaining viable. In order to establish a secondary site of growth, the cells must be capable of obtaining sustenance while floating freely in the blood stream, must overcome the host immunity and inflammatory mechanisms of destroying them, must lodge in the wall of a vessel, and must obtain a stoma at a foreign site.

Basically the change from a normal cell to a neoplastic one involves a change in cellular heredity. The daughter cells inherit their altered form and behavior pattern from their parent and pass the alteration to subsequent cells. Thus malignant cells are derived from previously normal ones. Carcinogenesis, the term applied to this process, involves the interaction of a previously unknown etiologic agent with susceptible tissue cells to produce the neoplastic change in form.

This condition has traditionally been treated using three types of methods: surgery, radiation and chemotherapy.

Surgery has been effective in certain cases, principally when the condition is discovered prior to metastasis. Radiation has also been effective to reduce and inhibit certain tumors. However, even with recent advances in diagnostic, surgical and radiation techniques, the incidences and numbers of deaths from this disease continue to increase.

Chemotherapy in the context of malignancy treatment involves the use of a chemical agent that will selectively 3,655,873 Patented Apr. 11, 1972 destroy cancer cells, leaving normal cells intact. Four types of agents are broadly known. These are cytotoxic, mutagenic chemicals (which are derivatives of nitrogen mustards); steroid hormones of androgenic and estrogenic activity; antimetabolites which are analogues of vitamins, purines and amino acids; and antibiotics derived from bacterial or plant somes. Specific agents of each group have found application in treatment of specific types of cancer, although permanent cures are rare.

These methods of treatment have accomplished, in some cases, amazing results based on new techniques and the developments derived through advanced research. This research, however, has not as yet isolated the etiologic factor by which metastasis develops, and therefore, the results of known treatments are not uniform.

Certain types of viruses have been identified as responsible for the change from a normal cell to a cancer cell in certain types of cancer. However, metastasis presents a different problem. It is known, for example, that certain types of cancer cells may be grown in a laboratory, and that complete surgical removal of a malignant tumor may result in a complete cure. Unfortunately even after complete surgical removal, the disease may recur at a later time in another area. Identification of the responsible virus does not explain the mechanism whereby the disease recurs, or fails to recur, after removal of apparently all diseased tissue, or the etiological factor which promotes recurrence in some individuals.

The method and preparation of this invention are directed to the treatment of several types of recurrent and metastatic cancer, to ease the pain of metastatic conditions, and to, at least temporarily, inhibit the spread of cancer cells. It cannot be established with certainty, as will subsequently be explained, the mechanism whereby the preparation achieves its beneficial results. However, the method and preparation stem from the discover of what appears to be an etiological factor in carcinoma. This factor may be a cause of the disease, or it may contribute to a symbiotic reaction with a related virus.

This invention was prompted by the discovery and isolation of Staphylococcus albus in cancer tissue. Cancer tissue segments were surgically removed under aseptic conditions and incubated in multiple electrolyte having a ten percent 10%) sugar concentration, and insulin at between 98-100 F. for about 48 hours. A smear of the cancer tissue was then placed on a blood agar plate and the plate incubated at 98.6 F. for 48 hours. This produced a luxurient growth of Staphylococcus albus.

As will be subsequently explained, the presence of Staphylococcus albus in metastatic cancer tissue was verified with repeated experiments. Identical techniques utilized with tissue from cancer free persons did not produce Staphylococcus cultures.

It is speculated that the presence of Staphylococcus albus is related to the physiologic, pathologic process which enters into the malignancy of the disease. If at the onset of carcinoma, the dermal or epithelial cells change from their normal inherited morphological process to become cells of phagocytic function, this change may be due to specific stimulation by a toxic agent from a germ or germs and virus. The phagocytic action on the part of the ectodermal cells is manifested in the evolution of their physiology. The cells depart from their basic pattern and move to various structures and organs of the body in pursuit of the toxic agent which has invaded the tissues and organs. However, cancer cells are incapable of destroying the agent, and therefore normal tissues are injured and destroyed by the conflict. During this process the cancer cells multiply and spread to various parts of the body in the phagocytic attempt to capture the agent. This agent may be Staphylococcus ulbus or it may be related to Staphylococcus albus, but the attemtp at phagocytosis is one explanation for malignancy, invasion and metastasis.

Another factor which may have an influence on malignancy involves immunity. It is well known that parent tumors may be completely removed with surgery. Then, in several months or years the cancer often will recur in the individual. In most cases recurrence will be by metastasis elsewhere in the body. Because cancer cells have been observed in circulation in the blood it is reasonable to speculate that the etiological agent, viral or bacterial, may be delayed in the process of metastasis due to an immune reaction on the part of the host. When this immunity is overcome, a recurrence and spreading of the disease takes place.

The process of this invention involves the culturing of Staphylococcus albus from metastatic carcinoma tissue, and incubation of this culture in a multiple electrolyte having ten percent sugar and insulin for approximately eight weeks to produce a solution having a distinctly therapeutic effect on metastatic carcinoma.

Accordingly it is an object of this invention to provide an agent isolated from culture of Staphylococcus albus taken from cancer tissue, which agent is effective to treat metastatic carcinoma.

It is another object to provide an agent effective to alleviate the metastatic condition in advance cases of cancer by inhibiting metastasis and relieving pain.

It is still another object to provide a Staphylococcus albus lysing solution by incubating cultures of said bacteria, derived from cancer tissue, in an electrolyte solution comprising ten percent invert sugar, salts, and insulin for approximately eight weeks at a temperature from 98- 100 F.

These and other objects will become readily apparent with reference to the following discussion.

The original discovery of the presence of Staphylococcus albus in cancer tissue came about through research directed to the study of the properties of cancerous cells. A segment of metastatic cancer of the abdomen, surgically removed from a human subject, was placed in normal saline solution and incubated at 98.4 F. for 48 hours. The segment was then smeared on a blood plate and incubated for seven days. Microscopic observation of the plate after the seven day incubation period revealed no bacterial growth.

Another segment of metastatic cancer tissue, surgically removed from the abdomen, was then placed in two ounces of an invert sugar electrolyte solution which also contained units of regular insulin. The tissue was incubated for 48 hours at 986. Following incubation, the tissue was smeared on a blood agar plate. The plate was then incubated at 986 for 48 hours. At the end of 48 hours a white uniform culture was observed to have formed on the plate. A slide was smeared with the culture, stained with methylene blue dye, and upon microscopic examination, the culture was observed to be Staphylococcus. To confirm that this culture was in fact a specie of Staphylococcus albus a smear of the culture was stained with gram stain, and found to be gram positive.

To confirm the presence of Staphylococcus albus in cancer tissue, this procedure was repeated with tissue segments obtained from 50 human patients. Cultures were taken from 20 patients suffering metastatic lesions of the abdomen, 23 suffering from cancer lesions of the breast, 2 from cancer of the ear, 1 having cancer of the thyroid gland, 1 advanced cancer of the stomach, 1 cancer of the esophagus, 1 cancer of the ovary, and 1 cancer of the rectum.

Forty-seven of these cultures incubated as described above produced pure cultures of Staphylococcus albus.

Furthermore, a segment of normal tissue from a breast in which a solitary cancer nodule was found was also taken, cultured as with the segments of cancer tissue and also produced a growth of Staphylococcus albus.

This procedure was repeated and confirmed with a ses= 0nd patient having a cancerous breast with multiple cancerous lesions. A segment from the apparently normal breast tissue was cultured and again produced Staphylococcus albus.

In a third case, in which a cancer on the periphery of the breast was diagnosed, two areas of the normal breast were cultured, and both areas also contained Staphylococcus albus.

In an effort to establish that the Staphylococcus albus from the tissue cultures was not, in fact, a contamination, the procedure was then repeated with twelve apparently normal cases. None of these cases utilized as a. control had diagnosed cancer present. In cultures surgically taken from a variety of different areas, the tissues, cultured as before, did .not show the presence of Staphylococcus albus. No tissue taken according to the identical techniques utilized with cancer patients produced Staphylococcus albus from patients not having cancer.

A killed suspension of the Staphylococcus albus cultured from cancerous tissues was then the subject of dermal tests on three disease free individuals. These tests produced no dermal reactions. The same tests were applied to cancer patients, and they also showed no protein reaction to the killed suspension.

In an effort to determine whether Staphylococcus albus alone was etiological in the cause of carcinoma, six rabbits and six hamsters were inoculated with pure cultures of Staphylococcus albus. There laboratory animals were observed for nine months and produced no reaction from the inoculations, even though a culture of the exudate in one hamster produced a pure culture of Staphylococcus albus. A suspension of live germs was injected in the mucocutaneous areas of the eyes, the mouth, and the rectum of these animals. The inability to produce a cancer reaction with Staphylococcus albus may be explained in dicate that an agent does in fact permeate the body prior that the germ may act in symbiosis with a related virus to produce cancer, or the inability to produce cancer may be due to an immune reaction in the animals.

Further research showed that no culture of Staphylococcus albus could be consistently obtained from cancer tissue by direct culture on 'blood plates. However, when the tissue was incubated in the electrolyte solution, a pure culture uniformly was obtained. If the Staphylococcus is a mutation from a virus stage, the electrolyte solution apparently offers a medium by which the virus stage may be advanced to the Staphylococcus albus cycle. This would further explain why Staphylococcus albus was successfully cultured from non-cancerous areas of a breast with cancer lesions. If there is a connection between the metastatic condition of cancer, and as yet an unidentified virus, the experiments with non-cancerous breast tissue will into surgical removal of a malignant tumor. This would explain why radical surgery is often ineffective in curing a patient of cancer.

However, the spread of the etiological agent throughout the body of a person suffering from cancer, does not achieve metastasis immediately, and this delay further suggests some type of immune reaction in the body.

The electrolyte utilized to culture cancerous tissue and the Staphylococcus albus developed therefrom, is commercially available and was used in an attempt to duplicate the natural system of electrolyte circulating in the body. The electrolyte is commercially obtainable from the McG-aw Laboratories, Inc, of Glendale, Calif. The electrolyte is marketed under the trade name Multiple Electrolyte 2 with 10% invert sugar. The concentration of the electrolyte, in microequivalents, per liter, is as follows:

Sodium 58 Potassium 25 Magnesium 6 Chloride 51 Lactate 25 Phosphate 13 The electrolyte is sterile and non-pyrogenic, and is an aqueous solution of the following chemical compounds in grams per 100 ml. of solution, or parts by weight per 100 parts water.

Invert sugar Sodium lactate 0.28 Sodium chloride 0.16 Potassium chloride 0.10 Dibasic potassium phosphate 0.10 Magnesium chloride hexahydrate 0.06 Sodium bisulfite less than 0.04 Monobasic sodium phosphate monohydrate 0.015

To this solution was added a quanity of regular insulin. The insulin was added because of the high quantity of sugar present in the electrolyte. The insulin was used solely to metabolize the sugar.

In a further effort to study the question of whether the Staphylococcus albus was a mutant stage in the etiology of carcinoma or in symbiosis with a virus of the same nature, a tissue from which Staphylococcus albus had originally been cultured was incubated in the electrolyte solution for about two months. The solution was then dropped on a blood plate, and the plate incubated for 48 hours. On examination it was noted that the middle zone of the plate contained no culture, while disposed around the rim of the plate, were a number of Staphylococcus colonies.

In an effort to evaluate this phenomenon, a 1,000 cc. electrolyte solution with 4,000 units of regular insulin was was utilized under aseptic conditions to incubate four blood plate cultures of Staphylococcus albus originally cultured from cancer tissue. The incubation period proceeded as follows: The temperature was maintained at 98- 100 F. After two days the container was agitated and 0.25 cc. withdrawn and cultured on a blood plate. After forty-eight hours a pure culture of Staphylococcus albus was observed. This procedure was repeated once weekly until at the end of the eighth week the blood plate on which the sample solution was cultured exhibited, after incubation, only two to three colonies of Staphylococcus albus.

From these experiments it was concluded that incubation of the cancer tissue in the electrolyte solution with insulin for about eight Weeks produced a lysing solution which may in fact be a bacteriophage active against Staphylococcus albus.

Toxicity of this lysing solution was evaluated in tests with rabbits. Several rabbits were intravenously injected with 4 cc. of the solution each. The observation period lasted one month following injection. During this period the rabbits exhibited no reaction or illness, and remained healthy and active.

The solution described above was then administered to 13 human cancer patients to evaluate whether lysing Staphylococcus albus Would have a beneficial elfect on metastatic carcinoma. The patients had advanced metastasis, and the majority were diagnosed as terminal prior to treatment.

In summary, the solution of this invention has been administered by injection in 2 cc. doses. The solution is nontoxic as shown in experiments with animals and has been administered with great success to thirteen human subjects. Of the thirteen, two made a complete recovery from metastatic cancer. Five showed a prolongation of life from six months to two years, and six showed temporary improvement. All ceased having pain and developed a good appetite upon treatment. None of the patients tested developed anemia until less than two to three months before death. Patients initially improved and were able to do ordinary work and drive automobiles. However, the improvement was not permanent in advanced cases.

In all patients a partial regression of the tumors during the time of improvement was observed, and in all patients a complete relief of pain due to the cancerous growth was also observed.

The solution of this invention is prepared by incubating a segment of cancer tissue for about 48 hours in 2 ounces of a 10% invert sugar electrolyte solution including 20 units regular insulin. The tissue is incubated at 98-99 F. Following incubation, the cancer tissue is smeared on a blood agar plate, and the plate is incubated for about 48 hours. At the end of 48 hours, a generous growth of Staphylococcus albus will be present on the plate. This may be confirmed through microscopic examination of a slide made from the growth and stained with methylene blue dye.

A suspension of Staphylococcus albus from the blood agar plate is then prepared in 10 cc. of the electrolyte solution. The suspension is incubated for 24 hours. Following incubation, 12 blood agar plates are seeded from the suspension. These 12 plates are then incubated at 98 to 99 F. for 2 to 3 days. The Staphylococcus albus cultured on the 12 plates is washed into a sterile container using the electrolyte as a wash solution. A large syringe, having 10 or 20 cc. capacity with a 20 gauge needle, is used to transfer the suspension of Staphylococcus albus from the container to a 1,000 cc. bottle containing the 10% invert sugar electrolyte solution and including 4,000 units of regular insulin. This solution is then incubated at 98-99 F. for from 6 to 8 weeks until a culture prepared from the solution exhibits no gowth. Following this incubation period, the solution is ready for use.

The incubation period is determined by culturing the solution weekly. After from 6 to 8 weeks, a culture prepared from the solution on a blood plate and incubated lfor 48 hours will exhibit no growth. When the weekly culture exhibits no growth the solution has reached the desired potency for treatment of cancer patients.

The solution is administered in doses of 2 cc. daily for the first four or five days. The solution is preferably administered by intramuscular injection. Following the initial pattern of daily injections the dosage is administered every 2 or 3 days for from 1 to 3 months.

Although the solution of this invention has not achieved a complete cure in all test cases and, in fact, is not claimed as a cure, in all cases administration caused an end to pain and a regression of tumor growth and metastasis. Even though the most advanced cases eventually succumbed from the disease, the solution of this invention exhibited a definite beneficial effect during usage and for periods when administration was interrupted.

Unfortunately, the beneficial tumor regression result in advanced cases was temporary, but relief from pain was definite and prolonged. However, in less advanced cases of metastasis, substantial prolongation of life resuited, and in cases having cancerous growth with only slight metastasis, complete regression of the tumor was achieved.

Although the exact mechanism whereby the solution of this invention achieves its beneficial result in metastatic carcinoma is not known, it may stimulate or duplicate an immunological response in the body interrupting the etiologic agent responsible for metastasis. Evidence suggests that the solution is a bacteriophage specific for Staphylococcus albus which organism was proven to be associated with metastatic carcinoma.

What is desired to be secured by US. Letters Patent is claimed as follows:

1. The method of preparing a therapeutic composition for use in treatment of metastatic carcinoma to induce tumor regression and to alleviate pain comprising the steps of: selecting a segment of cancerous tissue; incubating said tissue for approximately 48 hours at between 98 and 100 F. in 2 ounces of a multiple electrolyte solution containing 10% 'by weight invert sugar and 20 units of regular insulin; preparing a culture on a blood agar plate by smearing said incubated tissue on said plate; incubating said plate for 48. hours to form a bacterial culture on said plate; preparing a first suspension of said culture in 10 cc. of a multiple electrolyte solution; incubating said suspension for 24 hours; seeding a plurality of blood agar plates from the suspension; incubating said plates at 98 to 99 F. from 2 to 3 days to form a bacterial culture on each of said plates; washing said culture from said plates with a multiple electrolyte solution and collecting said wash as a second culture suspension in a sterile container; adding said second suspension to a multiple electrolyte solution containing 10% invert sugar and regular insulin; incubating said solution at 98 to 99 F. for from 6 to 8 Weeks until a culture prepared from said solution exhibits no growth.

2. The method of claim 1 wherein 12 blood agar plates are seeded from said first suspension and said culture obtained from said plates is added to 1,000 cc. of a multiple electrolyte containing 10% invert sugar and 4,000 units regular insulin.

3. The therapeutic composition obtained by the process of claim 1 useful to inhibit metastasis in carcinoma and to relieve pain associated with the disease in humans.

4. The method of claim 1 wherein said electrolyte comprises an aqueous solution in approximate parts by Weight of: 10 parts sugar; 0.28 part sodium lactate; 0.16 part sodium chloride; 0.10 part potassium chloride; 0.1 0 part dibasic potassium phosphate; 0.06 part magnesium chloride hexahydrate; less than 0.04 part sodium bisulfite; 0.015 part monobasic sodium phosphate monohydrate and 100 parts water.

5. The method of claim 4 wherein the culture prepared on said blood plate from said incubated cancerous tissue is Staphylococcus albus.

6. The method of treating metastatic malignancy to inhibit and repress tumor growth and to relieve the associated pain comprising the administration of about 2 cc. of the composition produced by the method of claim 1.

7. The method of claim 6 wherein the method of administration comprises injecting intramuscularly 2 cc. of said solution daily for from 4 to 5 days, and subsequently injecting 2 cc. of said solution every 2 to 3 days for from 1-3 months.

No references cited.

RICHARD L. HUFF, Primary Examiner UNITED STATES PATIENT owner CERTIFICATE Cl CORRECTION Patent NO. 3,655,873 Dated April 11, 1972 Inventor(s) Francis Ma Duffy It is certified that error appears in the above-identified patent 1 and that said Letters Patent are hereby corrected as shown below:

In Column 2, line 72, "attemtp" should read --attempt-- In Column 4, line 28, "There" should read --These--,-; line 36, "dicate that an agent does in fact permeate the body prior" should be canceled; line 52 after "in-" and before "to" in line 53, the following should be inserted --dicate that an agent does in fact permeate the body prior-=0 In Column 5, line 32, "was utilized" should read --utilized-- In Column 6, line 34, "lfor" should read "for".

Signed and sealed this 19th day of September 1972.

(SEAL) Attest:

EDWARD M.FLETCHER.JR.' ROBERT GUTTSCHALK Atte sting Officer Commissioner of Patents FORM PO-1050 (10-69) US OMM Dc some p69 a 0.5. eovsmmsm' rnmnuo OFFICE I969 0-366-334 

